EP0760393B1 - Procédé de chromatographie - Google Patents

Procédé de chromatographie Download PDF

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Publication number
EP0760393B1
EP0760393B1 EP96112637A EP96112637A EP0760393B1 EP 0760393 B1 EP0760393 B1 EP 0760393B1 EP 96112637 A EP96112637 A EP 96112637A EP 96112637 A EP96112637 A EP 96112637A EP 0760393 B1 EP0760393 B1 EP 0760393B1
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EP
European Patent Office
Prior art keywords
process according
fatty acids
mixture
acid
bar
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Lifetime
Application number
EP96112637A
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German (de)
English (en)
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EP0760393A3 (fr
EP0760393A2 (fr
Inventor
Gerd Brunner
Frank Reichmann
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DSM IP Assets BV
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F Hoffmann La Roche AG
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Publication of EP0760393A3 publication Critical patent/EP0760393A3/fr
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Classifications

    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
    • C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
    • C11C1/005—Splitting up mixtures of fatty acids into their constituents
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/40—Selective adsorption, e.g. chromatography characterised by the separation mechanism using supercritical fluid as mobile phase or eluent
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C07C51/47—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C67/00—Preparation of carboxylic acid esters
    • C07C67/48—Separation; Purification; Stabilisation; Use of additives
    • C07C67/56—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids

Definitions

  • the present invention relates to a method for obtaining unsaturated, optionally derivatized fatty acids from mixtures of these with other components, e.g. saturated, if necessary derivatized fatty acids, by column chromatography.
  • Eicosapentaenoic acid (“EPA”) and docosahexaenoic acid (“DHA”) are useful as active ingredients in the treatment and prevention of "lifestyle diseases", especially cardiovascular diseases, e.g. Heart attack and high cholesterol.
  • EPA Eicosapentaenoic acid
  • DHA docosahexaenoic acid
  • ⁇ -3 and ⁇ -6 PUFAs (with the first Double bond at the sixth carbon atom) plays an essential role in the Retinal, brain and general development of infants play. So ⁇ -3 and ⁇ -6 PUFAs have been detected in breast milk; it is considered ensured that breastfeeding children are significantly faster develop as comparable children who are not nursed.
  • eicosapentaenoic acid and docosahexaenoic acid in particular their positive effects in combating arteriosclerosis, the Lowering cholesterol and blood lipid levels and preventing Platelet aggregations, chronic inflammation, such as for example rheumatoid arthritis and neurodermatitis, and allergies currently under investigation.
  • Natural fats and oils, especially fish, marine and vegetable Oils are important sources of the ⁇ -3 and ⁇ -6 PUFAs, which are mainly found in Form of their glycerides and phospholipids occur and by many unwanted by-products and impurities are accompanied.
  • the ⁇ -3-PUFAs mentioned above eicosapentaenoic acid and docosahexaenoic acid are mainly contained in fish oils, while for example the ⁇ -6 PUFAs Arachidonic acid and linoleic acid especially in animal fats or vegetable oils, e.g. Corn oil.
  • the corresponding alkyl esters are obtained by alkaline hydrolysis of the mixtures containing the PUFAs, for example the fish, marine or vegetable oils themselves or refined forms thereof, and subsequent transesterification with alcohols, in particular with ethanol.
  • the subsequent technical separation of the alkyl ester mixture can be carried out, inter alia, by countercurrent extraction with a supercritical gas, in particular carbon dioxide, it being possible to carry out extensive separation according to the number of carbon atoms.
  • a supercritical gas in particular carbon dioxide
  • An SFC column chromatography is from the European patent publication (EP) 558.974 Process for obtaining unsaturated Fatty acids or derivatives thereof from plant-containing plants or animal mixtures.
  • EP European patent publication
  • stationary phase uses a defined phase.
  • the stationary phase is usually a silica gel or Alumina existing framework, which is mandatory with a Occupancy phase is provided, which free electron pairs and / or Has multiple bonds, such as in one Amino group (e.g. in aminopropyl) or a nitro group or one Phenyl group or a cyano group (e.g. in cyanopropyl) occurrence.
  • That effective separation of the individual PUFAs through SFC column chromatography with carbon dioxide as the mobile phase, however using an unoccupied framework, in particular Aluminum oxide that is reached does not come from EP 558.974 in any way out.
  • the object of the present invention was to develop a novel method to obtain valuable unsaturated fatty acids, as such or as derivatives, especially pre-prepared lower alkyl esters these fatty acids, from a mixture of the fatty acids and / or fatty acid esters to provide by column chromatography a process that largely does not have the disadvantages of the prior art having.
  • the method according to the invention is a Process for obtaining unsaturated fatty acids with at least sixteen carbon atoms in the molecule or a derivative thereof Fatty acid from a mixture of fatty acids and / or fatty acid derivatives by column chromatography with supercritical or liquid carbon dioxide as a mobile phase, which is characterized in that as Alumina pretreated with alkali is used.
  • the method according to the invention is carried out in such a way that that the desired mixture of Fatty acids and / or fatty acid derivatives with the mobile phase of brings supercritical or liquid carbon dioxide together, the whole thing possibly followed by another mobile phase, on the one mentioned above stationary phase packed chromatography column there and then can flow through (eluting), the elution among the selected Temperature and pressure conditions occur and due to the strong Interactions between the stationary phase and the individual Components of the mixture a temporal separation of these components is achieved, which elutes one after the other from the column, in carbon dioxide dissolved components (eluate) after sequential detection (determination) in collecting receptacles determined by the detection means and that Carbon dioxide from the respective groupage by decompression (Volatilization) removed, so that the received, separated components or "fractions" - (including the desired ones unsaturated fatty acids or fatty acid derivatives) free of carbon dioxide in the individual receptacles. If desired, after the Elute and exit the column the eluate one or
  • oils or fats of vegetable or animal (including marine) origin can be used, which include the desired fatty acids in the form of triglycerides, amides, phospholipids, lactones and salts, and other other fatty acids and derivatives such as triglycerides etc., thereof, sterols , e.g. cholesterol, vitamins, e.g. tocopherols, and materials that are normally regarded as impurities, e.g.
  • PCB polychlorobiphenyl
  • PAH polyaromatic hydrocarbons
  • pesticides pesticides
  • dioxins heavy metals
  • heavy metals oxidation and decomposition products of saturated fatty acids or their derivatives, from previously contain any remaining solvents or reagents, etc.
  • a raw material oil or fat which may have been refined and chemically treated, esterified or transesterified beforehand is used.
  • the corresponding, optionally previously refined oil or fat is subjected to acidic or basic hydrolysis, the triglycerides contained therein being converted into the corresponding acids, and esterified with a C 1-6 -alkyl alcohol, preferably with ethanol, the triglycerides being the unsaturated and saturated fatty acids are transesterified into the corresponding lower alkyl esters.
  • the hydrolysis or esterification can be carried out in a conventional manner.
  • Preferred raw materials, which may have been refined and / or chemically treated beforehand, are fish oils, e.g.
  • sardine and tuna oil since they are valuable sources of the highly sought-after eicosapentaenoic acid and docosahexaenoic acid, as well as animal fats and vegetable oils, e.g. corn oil, since these are in turn Valuable sources of arachidonic acid and linoleic acid, which are also in demand, are especially fish oils.
  • the mixture of fatty acids and / or fatty acid esters normally without dilution together with the supercritical or liquid carbon dioxide to those used with the invention given stationary phase packed chromatography column can however in advance in a suitable solvent, e.g. one lower alkane, preferably n-hexane, are dissolved.
  • a suitable solvent e.g. one lower alkane, preferably n-hexane
  • the mixture is used undiluted.
  • Carbon dioxide is known to be in the form of Carbon dioxide, which is at a temperature of at least about 31 ° C and at a pressure of at least about 73 bar is maintained and neither pure liquid still pure gaseous, but a hybrid of the two physical Forms is. That which can be used as an alternative in the process according to the invention liquid carbon dioxide has a temperature of less than about 31 ° C and a pressure which is above about 73 bar.
  • the advantages of using carbon dioxide are its Non-toxicity, non-flammability and easy removal through Decompress the collected eluates without a potential harmful residue with the separated unsaturated fatty acid or with a derivative of it.
  • the carbon dioxide is high Purity and inexpensive widely available and can, if desired with an organic cosolvent ("modifier"), e.g. the one above mentioned n-hexane, but also a lower alkanol, for example Methanol or ethanol, and a lower aliphatic ketone, e.g. Use acetone as part of the mobile phase.
  • modifier organic cosolvent
  • the critical temperature of carbon dioxide is not much higher than room temperature and that Unsaturated fatty acids to be obtained according to the invention or Fatty acid derivatives are temperature sensitive (thermolabile) is suitable
  • carbon dioxide is also excellent than in Mobile phase used according to the method of the invention.
  • the aluminum oxide used in the process according to the invention as the stationary phase - and characteristic of the invention - is expediently present as homogeneously packed, non-uniform or, preferably, spherical (spherical) particles, the particle size of which is approximately 5 to 25 ⁇ m.
  • spherical alumina is readily available commercially.
  • Aluspher®Al and Spherisorb Alumina may be mentioned as examples of commercially available aluminum oxides; the former has a specific surface area S BET of 170 m 2 / g, a pore volume Vp of 0.5 ml / g, a pore diameter D of 100 ⁇ and a particle size dp 50 of 5 ⁇ m, while the latter has an S BET of 93 m 2 / g, a D of 130 ⁇ and a dp 50 of also 5 ⁇ m.
  • Broken materials are offered by various manufacturers, for example ICN Biomedicabs, Inc. These aluminum oxides have particle size distributions of 3-6 ⁇ m, 7-12 ⁇ m, 10-18 ⁇ m or 18-32 ⁇ m.
  • the specific surface S BET of these materials is 200 m 2 / g; the pore volume and pore diameter are in the range of spherical materials.
  • the alumina is before Subjected to use of an alkaline pretreatment. at this pretreatment is basically the contact of the particulate alumina with an aqueous solution of a Alkali metal or alkaline earth metal hydroxide, such as sodium, Potassium or calcium hydroxide, in the pH range from about 10 to about 13 several hours, suitably over about 8 to about 20 hours.
  • a Alkali metal or alkaline earth metal hydroxide such as sodium, Potassium or calcium hydroxide
  • the Concentration of the aqueous alkali metal or alkaline earth metal hydroxide solution expediently lies in the range from approximately 0.01 M to approximately 0.1 M.
  • the aqueous alkali metal or alkaline earth metal hydroxide solution with a water-miscible or water-soluble organic solvents are added to the viscosity of the solution to set a value that is workable from a technical point of view. This will for example with the polar organic solvent acetonitrile or Acetone reached.
  • the 10:90 (V / V) mixture has proven particularly suitable of acetonitrile and 0.1 M aqueous sodium hydroxide solution, which has a pH of about 13.
  • the alkaline pretreatment of the Aluminum oxide expediently by continuous flow the alkaline solution through that already in the chromatography column packed alumina over several hours, preferably about 12 to 16 hours followed by rinsing the pretreated so Alumina with distilled water until the escaping rinses are neutral.
  • the column for about 4-10 Hours between 50 and 90 ° C and then the pretreated Aluminum oxide with a lower alkane, preferably rinse n-heptane for several hours, preferably about 8 to 20 hours.
  • the pretreated aluminum oxide as a stationary phase in the method according to the invention can be used.
  • the use of the Alumina pretreated with alkali is a preferred aspect of the inventive method.
  • the carbon dioxide used as the mobile phase in the process according to the invention In order for the carbon dioxide used as the mobile phase in the process according to the invention to be maintained in the supercritical or liquid range, certain temperature and pressure conditions must be maintained, not only when the carbon dioxide is introduced into the chromatography column packed with the stationary phase, but also during the subsequent elution.
  • the process is expediently carried out in the temperature range from approximately 30 ° C. to approximately 100 ° C. and at a pressure of approximately 140 bar to approximately 320 bar, the suitable pressure being lower under otherwise identical process conditions when the aluminum oxide pretreated with alkali is used instead of the untreated aluminum oxide is.
  • the temperature range is preferably approximately 45 ° C. to approximately 75 ° C. and the relevant pressure range is approximately 220 bar to approximately 260 bar.
  • the density of carbon dioxide can be adjusted via pressure and temperature, and in the last-mentioned temperature and pressure range there is an optimal carbon dioxide density of about 720 kg / m 3 to about 850 kg / m 3 .
  • Aluminum oxide acts as a stationary phase during the implementation of the method according to the invention the chromatographic separation the various unsaturated fatty acids or fatty acid derivatives and, if present, saturated fatty acids or fatty acid derivatives, namely on the one hand according to the chain length (number of carbon atoms) and on the other hand according to the degree of saturation (number of double bonds: the higher these Number, the lower the degree of saturation). From a certain number Double bonds seem to increase the selectivity regarding the degree of saturation predominate.
  • fatty acids or fatty acid derivatives to be separated via only one double bond, such as the Erucic acid (22 carbon atoms, 1 double bond: "22: 1"), so the selectivity occurs regarding the chain length clearly in the background, and the fatty acids or derivatives elute before the range of shorter, but more unsaturated Fatty acids or derivatives, such as the 20: 5 fatty acid eicosapentaenoic acid.
  • the order of the eluate changes according to this Treatment is not, but the tailing of the polar is reduced polyunsaturated fatty acids, and the elution times are reduced clear.
  • the elution is carried out with the same Temperature and pressure conditions on the alkaline treated Alumina after almost a third of the time and with strong reduced tailing. For this reason, with Alumina pretreated with alkali in the invention Process used.
  • Detection on the chromatography column in succession eluting constituents dissolved in carbon dioxide (eluates) are carried out sequentially preferably via a UV detector, followed by a flame ionization detector (FID).
  • the UV detector enables an assessment of the Components according to the number of double bonds and can therefore be used limited structural elucidation of individual components.
  • Under Use of the FID will allocate the various eluates to the Collecting containers electronically.
  • Such technology is in itself known, as well as the way in which the carbon dioxide (by Decompression) is removed from the respective groupage.
  • the method according to the invention is very preferably suitable for Obtaining the 20: 5 fatty acid eicosapentaenoic acid and the 22: 6 fatty acid Docosahexaenoic acid, either directly or through a derivative thereof, preferably the ethyl ester.
  • the equipment is continuously supplied with a mobile phase via a carbon dioxide (CO 2 ) bottle.
  • CO 2 carbon dioxide
  • the mobile phase can be operated in the supercritical range (with CO 2 above approx. 31 ° C and 73 bar) or in the subcritical range.
  • the CO 2 is drawn from the bottle into the chamber of the pump, where it is first condensed to -6 ° C using a thermostat.
  • the piston is driven by a stepper motor and thus compresses the mobile phase to a desired pressure.
  • the pressure of the syringe pump and the temperature of the column oven can be set by computer.
  • the mobile phase flows through the apparatus at the selected pressure and first passes through the injection system.
  • the sample is fed into the mobile phase flowing through a two-way valve.
  • the task time can be varied between 0 and 999 msec.
  • the sample volume is a constant 0.5 ⁇ l.
  • the sample then reaches the column with the mobile phase.
  • the mixture is separated due to the strong interactions between the stationary phase and the individual components of the sample solution.
  • the individual constituents of the mixture are (ideally) eluted from the column in succession. Since the column is in an oven, it is also possible to influence the elution behavior via the temperature.
  • the detection (determination) of the substances separated on the column is carried out sequentially using a UV detector and then using a flame ionization detector (FID).
  • a chromatography column packed with alumina is passed through rinsing for several hours (12-24 hours) with a mixture of 0.1 M treated aqueous sodium hydroxide solution in acetonitrile (90: 10% v / v). The pH of the solution is therefore 13.
  • the column is distilled Washed water neutral, for about 4-10 hours between 50 and 90 ° C heated and then reconditioned for 12-18 hours with n-heptane.
  • An apparatus from New Ways of Analytics (see Figure 4) is used to operate columns with a larger inner diameter and the associated increase in throughput.
  • the equipment is continuously supplied with a mobile phase via a CO 2 feed line.
  • the mobile phase can be operated in the supercritical range (with CO 2 above approx. 31 ° C and 73 bar) or in the subcritical range.
  • the CO 2 that flows from a CO 2 container into the thermostats of the PM-101 pressure module is condensed to -6 ° C by means of a thermostat.
  • a pneumatic piston pump compresses the mobile phase to pressures between 300-400 bar.
  • a downstream pressure reduction module PR-102 the compressed mobile phase is reduced to the desired pressure and additional pressure fluctuations, which result from the functioning of the pressure module PM-101, are damped.
  • the mobile phase flows through the apparatus at the pressure selected in the PR-102 pressure reduction module and first passes through the injection valve.
  • the sample is fed into the mobile phase flowing past via a six-way valve (Rheodyne 7125).
  • the sample feed volume can be varied from 5 ⁇ l to several milliliters by selecting appropriate test loops.
  • the sample then reaches the column with the mobile phase.
  • the mixture is separated due to the strong interactions between the stationary phase and the individual components of the sample solution. Ideally, the individual components of the mixture are eluted one after the other.
  • the column Since the column is in an oven, it is also possible to influence the elution behavior via the temperature.
  • the substances separated on the column are recorded by a UV detector and from there transmitted to a chromatography software.
  • the software enables the switching of valves that enable the separated components to be removed.
  • the relaxation of the supercritical phase is finally carried out via the relaxation unit PE-103.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Fats And Perfumes (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Steroid Compounds (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)

Claims (10)

  1. Procédé destiné à obtenir des acides gras insaturés, à au moins seize atomes de carbone dans la molécule, ou leurs C1-6-esters d'alkyle à partir d'un mélange d'acides gras et/ou de dérivés d'acides gras grâce à une chromatographie sur colonne avec du dioxyde de carbone liquide ou supercritique comme phase mobile, caractérisé en ce qu'on sépare les acides gras sous forme de C1-6-ester d'alkyle sur de l'oxyde d'aluminium traité au préalable aux alcalis comme phase stationnaire et si souhaité, l'ester obtenu se saponifie en acides libres.
  2. Procédé selon la revendication 1, caractérisé en ce que l'oxyde d'aluminium traité au préalable aux alcalis se présente sous la forme de particules en forme de billes compactées de façon homogène dont la taille de particule s'élève à environ 5 à 25 µm.
  3. Procédé selon la revendication 1 ou 2, caractérisé en ce que le traitement préalable de la phase stationnaire a lieu par le contact de l'oxyde d'aluminium sous forme de particules avec une solution aqueuse d'un hydroxyde de métal alcalino-terreux ou de métal alcalin dans l'intervalle de pH d'environ 10 à environ 13 pendant environ 8 à environ 20 heures.
  4. Procédé selon la revendication 3, caractérisé en ce que pour le réglage de la viscosité de la solution aqueuse sur une valeur techniquement utilisable pour la travailler, la solution est complétée par un solvant organique soluble dans l'eau ou miscible dans l'eau, en particulier avec de l'acétonitrile ou de l'acétone.
  5. Procédé selon la revendication 4, caractérisé en ce que pour le traitement préalable de l'oxyde d'aluminium, un mélange à 10:90 (V/V) d'acétonitrile et 0,1 M d'une solution aqueuse d'hydroxide de sodium est utilisé.
  6. Procédé selon l'une quelconque des revendications 1 à 5, caractérisé en ce qu'on utilise une huile de poisson éventuellement préalablement traitée chimiquement et/ou raffinée comme mélange d'acides gras et/ou de dérivés d'acides gras.
  7. Procédé selon la revendication 6, caractérisé en ce qu'on utilise une huile de poisson préalablement esterisée avec un C1-6-alcanol, en particulier une huile de poisson estérisée avec de l'éthanol, comme mélange d'acides gras et/ou de dérivés d'acides gras.
  8. Procédé selon l'une quelconque des revendications 1 à 7, caractérisé en ce que le procédé est exécuté dans l'intervalle de température d'environ 30° C à environ 100° C, et sous une pression d'environ 140 bar à environ 320 bar.
  9. Procédé selon la revendication 8, caractérisé en ce que l'intervalle de température s'élève entre environ 45° C et environ 75° C, et l'intervalle de pression correspondant entre 220 bar et environ 260 bar.
  10. Procédé selon l'un quelconque des revendications 1 à 9, caractérisé en ce qu'il est utilisé pour obtenir un acide eicosapentaenoique, un acide docosapentaenoïque ou un acide docosahexaenoïque.
EP96112637A 1995-08-17 1996-08-05 Procédé de chromatographie Expired - Lifetime EP0760393B1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CH235595 1995-08-17
CH2355/95 1995-08-17
CH235595 1995-08-17

Publications (3)

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EP0760393A2 EP0760393A2 (fr) 1997-03-05
EP0760393A3 EP0760393A3 (fr) 1997-10-29
EP0760393B1 true EP0760393B1 (fr) 2003-06-04

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Country Status (8)

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US (1) US5777141A (fr)
EP (1) EP0760393B1 (fr)
JP (1) JP3739494B2 (fr)
CN (1) CN1137745C (fr)
AT (1) ATE242303T1 (fr)
DE (1) DE59610489D1 (fr)
DK (1) DK0760393T3 (fr)
ES (1) ES2200023T3 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7439267B2 (en) 2001-01-25 2008-10-21 Pfizer Italia S.R.L. Essential n-3 fatty acids in cardiac insufficiency and heart failure therapy

Families Citing this family (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE400358T1 (de) 1997-12-24 2008-07-15 Cepheid Vorrichtung und verfahren zur lyse
US7914994B2 (en) * 1998-12-24 2011-03-29 Cepheid Method for separating an analyte from a sample
DE60012934T2 (de) 1999-02-26 2005-08-25 Martek Biosciences Corp. Verfahren zum Abtrennen von einem Docosahexaensäure enthaltenden Triglycerid aus einem Triglyceridgemisch
AU4030600A (en) * 1999-03-26 2000-10-16 Martek Biosciences Corporation Specific binding assay for docosahexaenoic acid
DE19923558A1 (de) * 1999-05-21 2000-11-23 K D Pharma Bexbach Gmbh Verfahren zur Herstellung von geruchs- und geschmacksfreien ungesättigten Fettsäuren aus Naturölen und deren Verwendung
US6664104B2 (en) * 1999-06-25 2003-12-16 Cepheid Device incorporating a microfluidic chip for separating analyte from a sample
JP5503856B2 (ja) * 2008-09-10 2014-05-28 キユーピー株式会社 高度不飽和脂肪酸誘導体の取得方法
WO2011005505A2 (fr) 2009-06-22 2011-01-13 Johnson Matthey Public Limited Company Procédé pour la purification de prostaglandines
US20110033595A1 (en) * 2009-08-10 2011-02-10 Rudolf Krumbholz Fatty acid fractionation process, fatty acid products and use thereof
RU2538981C2 (ru) 2009-12-30 2015-01-10 Басф Фарма (Калланиш) Лимитед Способ хроматографического разделения с псевдодвижущимся слоем
GB201111591D0 (en) 2011-07-06 2011-08-24 Equateq Ltd Further new process
GB201111601D0 (en) 2011-07-06 2011-08-24 Equateq Ltd New process
GB201111589D0 (en) 2011-07-06 2011-08-24 Equateq Ltd New modified process
GB201111594D0 (en) 2011-07-06 2011-08-24 Equateq Ltd New improved process
GB201111595D0 (en) 2011-07-06 2011-08-24 Equateq Ltd Improved process
GB201300354D0 (en) 2013-01-09 2013-02-20 Basf Pharma Callanish Ltd Multi-step separation process
US9428711B2 (en) 2013-05-07 2016-08-30 Groupe Novasep Chromatographic process for the production of highly purified polyunsaturated fatty acids
US8802880B1 (en) 2013-05-07 2014-08-12 Group Novasep Chromatographic process for the production of highly purified polyunsaturated fatty acids
US10151733B2 (en) 2013-09-13 2018-12-11 Waters Technologies Corporation Catalytic oxidation of polar modifiers in chromatographic mobile phases
EP2883860B1 (fr) 2013-12-11 2016-08-24 Novasep Process Procédé chromatographique de production d'acides gras polyinsaturés
US10975031B2 (en) 2014-01-07 2021-04-13 Novasep Process Method for purifying aromatic amino acids
SG10201912618XA (en) * 2015-08-31 2020-02-27 Nippon Suisan Kaisha Ltd Free polyunsaturated fatty acid-containing composition and manufacturing method therefor

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE741359C (de) * 1937-09-08 1943-11-10 Dr Hans P Kaufmann Verfahren zur Fraktionierung von Gemischen aus Fettsaeuren u. dgl.
JPH0653702B2 (ja) * 1986-06-11 1994-07-20 エーザイ株式会社 ビタミンｋ▲下２▼異性体の分離精製法
US4880543A (en) * 1987-08-31 1989-11-14 Aluminum Company Of America Supercritical fluid chromatography packing material containing alumina
DE3929555A1 (de) * 1989-09-06 1991-03-07 Sueddeutsche Kalkstickstoff Verfahren zur entfernung von cholesterin bzw. cholesterinestern aus lebensmitteln
DE4206539A1 (de) * 1992-03-02 1993-09-09 K D Pharma Gmbh Verfahren zum gewinnen ungesaettigter fettsaeuren
ATE147776T1 (de) * 1993-04-29 1997-02-15 Norsk Hydro As Verfahren zur chromatografischer fraktionierung von fettsäuren und ihre derivaten

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7439267B2 (en) 2001-01-25 2008-10-21 Pfizer Italia S.R.L. Essential n-3 fatty acids in cardiac insufficiency and heart failure therapy

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EP0760393A3 (fr) 1997-10-29
ES2200023T3 (es) 2004-03-01
JP3739494B2 (ja) 2006-01-25
JPH09104894A (ja) 1997-04-22
DE59610489D1 (de) 2003-07-10
CN1137745C (zh) 2004-02-11
ATE242303T1 (de) 2003-06-15
CN1154868A (zh) 1997-07-23
US5777141A (en) 1998-07-07
EP0760393A2 (fr) 1997-03-05
DK0760393T3 (da) 2003-09-29

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